A Potent d-Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo

Uppalapati, Maruti; Lee, Dong Jun; Mandal, Kalyaneswar; Li, Hongyan; Miranda, Les P.; Lowitz, Joshua; Kenney, John; Adams, Jarrett J.; Ault-Riché, Dana; Kent, Stephen B. H.; Sidhu, Sachdev S.

doi:10.1021/acschembio.5b01006

Title: A Potent d-Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo

Journal Article · Wed Feb 03 00:00:00 EST 2016 · ACS Chemical Biology

DOI:https://doi.org/10.1021/acschembio.5b01006· OSTI ID:1236725

Uppalapati, Maruti ^[1]; Lee, Dong Jun ^[2]; Mandal, Kalyaneswar ^[2]; Li, Hongyan ^[3]; Miranda, Les P. ^[4]; Lowitz, Joshua ^[5]; Kenney, John ^[5]; Adams, Jarrett J. ^[1]; Ault-Riché, Dana ^[6]; Kent, Stephen B. H. ^[2]; Sidhu, Sachdev S. ^[1]

Banting and Best Department of Medical Research and Department of Molecular Genetics, the Donnelly Centre, University of Toronto, Toronto, Ontario, Canada M5S 3E1
Department of Chemistry, Department of Biochemistry and Molecular Biology, Institute for Biophysical Dynamics, University of Chicago, Chicago, Illinois 60637, United States
Pharmacokinetics &, Drug Metabolism, Amgen, Inc., Thousand Oaks, California 91320, United States
Therapeutic Discovery, Amgen, Inc., Thousand Oaks, California 91320, United States
Antibody Solutions, Sunnyvale, California 94089, United States
Reflexion Pharmaceuticals, Inc., San Francisco, California 94104, United States

Polypeptides composed entirely of d-amino acids and the achiral amino acid glycine (d-proteins) inherently have in vivo properties that are proposed to be near-optimal for a large molecule therapeutic agent. Specifically, d-proteins are resistant to degradation by proteases and are anticipated to be nonimmunogenic. Furthermore, d-proteins are manufactured chemically and can be engineered to have other desirable properties, such as improved stability, affinity, and pharmacokinetics. Thus, a well-designed d-protein therapeutic would likely have significant advantages over l-protein drugs. Toward the goal of developing d-protein therapeutics, we previously generated RFX001.D, a d-protein antagonist of natural vascular endothelial growth factor A (VEGF-A) that inhibited binding to its receptor. However, RFX001.D is unstable at physiological temperatures (T_m = 33 °C). Here, we describe RFX037.D, a variant of RFX001.D with extreme thermal stability (T_m > 95 °C), high affinity for VEGF-A (K_d = 6 nM), and improved receptor blocking. Comparison of the two enantiomeric forms of RFX037 revealed that the d-protein is more stable in mouse, monkey, and human plasma and has a longer half-life in vivo in mice. Significantly, RFX037.D was nonimmunogenic in mice, whereas the l-enantiomer generated a strong immune response. These results confirm the potential utility of synthetic d-proteins as alternatives to therapeutic antibodies.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1236725

Alternate ID(s):: OSTI ID: 1249238

Journal Information:: ACS Chemical Biology, Journal Name: ACS Chemical Biology Vol. 11 Journal Issue: 4; ISSN 1554-8929

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 62 works

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Title: A Potent d-Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo

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